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US 2021/0239710 A1Published application
Flow cytometry for monitoring histone h3 methylation status
AssigneeEpizyme, Inc.
InventorChristopher PLESCIA
Life of this patent
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Abstract
The present disclosure relates to methods of detecting histone epigenetic modifications and compositions comprising inhibitors of human histone methyltransferase EZH2 and their use for the treatment of cancer.
Claims29
A method comprising
- quantifying trimethylation of lysine 27 of histone H3,
- quantifying total H3 level, and
- immunophenotyping of a population of cells obtained from a subject.
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A method comprising:
- (a) contacting at least one cell in a sample from a subject with a permeabilizing composition to obtain a permeabilized sample;
- (b) contacting the permeabilized sample with a first fluorescent-conjugated antibody that specifically binds an epitope within H3K27me1, H3K27me2, or H3K27me3;
- (c) contacting the permeabilized sample with a second fluorescent-conjugated antibody that specifically binds an epitope within H3;
- (d) detecting a first fluorescent signal from the first fluorescent-conjugated antibody;
- (e) detecting a second fluorescent signal from the second-fluorescent conjugated antibody;
- and
- (f) determining an amount of H3 that is epigenetically-modified comprising dividing a value of the first fluorescent signal by a value of the second fluorescent signal, to quantify the epigenetic modification of the histone H3 in the sample.
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Dependencies are inferred from the claim text.